Detection of restensosis risk in patients receiving a stent

ABSTRACT

Provided is a method of selecting a stent for implantation in the circulatory system of a human being. The method comprises obtaining a blood sample from a patient who requires implantation of a stent and testing said blood sample to determine a platelet coagulability level. The determined platelet coagulability level of said blood sample is compared with a threshold level of blood platelet coagulability. A determined platelet coagulability level above said threshold level indicates that a risk of restenosis is relatively high. If the determined platelet coagulability level is below said threshold level, a bare metal stent is selected. If the determined platelet coagulability level is at or above said threshold level, a drug-eluting stent is selected.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.15/345,159, filed on Nov. 7, 2016, which is a continuation of Ser. No.14/943,939, filed on Nov. 17, 2015, which is a continuation of Ser. No.13/309,121 filed on Dec. 1, 2011, now U.S. Pat. No. 9,188,597, which isa continuation of Ser. No. 11/663,599, filed on Mar. 23, 2007, now U.S.Pat. No. 8,070,678, which is the National Stage of InternationalApplication No. PCT/US06/38303, filed on Oct. 4, 2006, which claims thebenefit of U.S. Provisional Application No. 60/723,453, filed on Oct. 5,2005. The applications to which the present application claims benefitare herein incorporated by reference in their entireties.

FIELD OF THE INVENTION

The present invention relates to assessing the risk of developingrestenosis in patients treated with stents with atherosclerotic vasculardisease and using the risk assessment to determine the intensity ofantiproliferative therapy administered to the vascular wall by means ofa drug-eluting stent.

BACKGROUND OF THE INVENTION

Restenosis remains a major limiting factor in the percutaneous treatmentof coronary artery disease. Despite improvements in restenosis ratesachieved from the use of stents and the introduction of drug-elutingstents, restenosis persists in a small percentage of patients. Thesequence of events contributing to restenosis is initiated at the stentsite as a result of arterial wall trauma, endothelial injury and therelease of growth factors, chemoattractants, and inflammatory mediators.These events induce platelet and leukocyte activation and trigger thecoagulation cascade. Although the underlying pathophysiology is notuniformly accepted, major pathological findings in acute or chronicstent failure include the deposition of fibrin and platelets, suggestingthat a key event in development of restenosis is thrombus formation.

Treatment with stents reduces restenosis compared to balloon angioplastyand now drug-eluting stents have further reduced restenosis rates.However, it is well recognized that most patients treated with baremetal stents develop clinically irrelevant degrees of intimalproliferation and therefore would not necessarily have benefited fromthe implantation of a drug-eluting stent. See R. Moreno, et al.,Drug-eluting stent thrombosis: results from a pooled analysis including10 randomized studies; J Am Coll Cardiol. 2005; 45: 9549. Nevertheless,the current practice is to implant the more costly drug-eluting stentsin the majority of patients undergoing percutaneous intervention. See H.C. Lowe, et al., Coronary in-stent restenosis: current status and futurestrategies; J Am Coll Cardiol. 2002; 39: 183-93. The ability to predictwhich patients are most prone to developing neointimal formation couldlead to more selective use of drug-eluting stents and tailor theintensity of antiproliferative therapy.

Platelet-related periprocedural thrombotic and inflammatory processesthat influence neointimal hyperplasia and angiogenesis are consideredimportant risk factors for restenosis. See B. Chandrasekar et al.,Platelets and restenosis; J Am Coll Cardiol. 2000; 35: 555-62. Inaddition, preexisting inflammatory mediators and hypercoagulable factorshave also been proposed to influence the process. Despite an establishedmechanism linking thrombogenesis to the restenosis process, there arefew data in humans that have examined the relation of ex vivomeasurements of platelet reactivity to restenosis. This informationmight be clinically useful in evaluating patients undergoingPercutaneous Cardiovascular Intervention (PCI) to identify a subgroupwho may benefit from more aggressive therapy aimed at disrupting thesequence of events leading to restenosis. The ability to predict whichpatients are most prone to developing neointimal formation could alsolead to more selective use of the more costly drug-eluting stents (DES).Use of DES has reduced restenosis rates. Currently, DES are routinelyimplanted in the majority of patients undergoing PCI withoutconsideration as to whether the clinically irrelevant degrees of intimalproliferation that develop in most patients treated with bare metalstents warrant this practice. Moreover, there is concern that DES have agreater risk of thrombosis than bare metal stents.

At this time, there is no uniformly accepted method to determine whichpatients are at greatest risk for developing stent restenosis. A majorcost savings would result from a method that reliably predicted thosepatients at greatest risk. These patients would be treated with the morecostly drug-eluting strategy whereas those at minimal risk would receivethe less expensive bare metal stent. It is well known that specificangiographic and clinical features are associated with a higher risk ofrestenosis. These include the presence of diabetes, small vessels, longlesions and bifurcation disease. In addition, a strategy that determineswho will benefit from DES will entail much less use of dual antiplatelettherapy that is required indefinitely in patients treated with DES dueto the excess hazard of stent thrombosis. Presently, there are nolaboratory tests that predict the occurrence of restenosis.

Platelets play a fundamental role in the genesis of stent restenosis bymodulating coagulation, inflammation, and smooth muscle proliferation.Thrombi with high tensile strength may facilitate neointimal hyperplasiaat the stent site. Platelet-related periprocedural thrombotic andinflammatory processes that influence neointimal hyperplasia andangiogenesis are considered important risk factors for restenosis inanimal models. See P. A. Gurbel, et al., Platelet reactivity in patientsand recurrent events post-stenting: results of the PREPARE POST-STENTINGStudy; J Am Coll Cardiol. 2005; 46: 1820-26. Despite these establishedmechanisms, there are few data in humans that examined the relation ofex vivo measurements of platelet reactivity to restenosis. Moreover,preexisting inflammatory and hypercoaguable factors have also beenproposed as important factors influencing restenosis. Moreover, patientswith rapid thrombin generation would be expected to readily formthrombi.

There is a need in the field for a method to accurately risk-stratifypatients for restenosis. This methodology would therefore tailor patienttherapy during stent implantation. This method would assist in thedecision making for using a bare metal stent versus a drug-eluting stentand, moreover, would also determine the intensity of drug delivery basedon the individual patient's risk.

SUMMARY OF THE INVENTION

The invention features methods and compositions for assessing the riskof developing restenosis in patients with vascular disease undergoingstenting. The invention is based on the discovery that a plateletmediated hypercoaguable state is an important risk factor for thedevelopment of restenosis and identifies patients with the highest riskof needing a subsequent revascularization procedure. Accompanying theplatelet-mediated hypercoaguable state is the presence of rapidfibrin-platelet clot formation which is a marker of the speed andintensity of thrombin generation; and a strong platelet-fibrin clot(i.e. high tensile strength). Therefore, any tool that can measureplatelet-mediated hypercoaguability would be expected to predictrestenosis. In my experiments, I measured these properties (i.e., thespeed of thrombin generation by the parameter, R, and the strength ofthe platelet-fibrin clot by the parameter, MA) by thrombelastography,but other methods including enzyme linked immunosorbent assays tomeasure thrombin generation and devices to measure platelet reactivityincluding aggregometers, and flow cytometry and other tools that measurethe viscoelastic properties of the clot would be expected to predictrestenosis.

In an embodiment of particular interest, the risk of developingrestenosis is assessed by determining the maximum tensile strength ofthe clot formed in the blood of the particular patient after stimulatingthe blood with an agonist that generates thrombin. The restenosis scoreis then measured and the individual risk is assessed based on therelation of the score to a chosen threshold level. The risk level isthen used as a guide to determine whether to treat the patient with abare metal stent as compared to a drug-eluting stent. In those patientswith the highest risk, the most intensive drug delivery is chosen.

The invention is advantageous in that, prior to the invention, noreadily available or accepted methodology was available to assess therisk for restenosis of the individual patient undergoing coronarystenting. Current practice is to treat patients with drug-eluting stentsirrespective of an assessment of their risk for restenosis. Drug-elutingstents are expensive and require prolonged therapy with expensiveantiplatelet agents. Many patients are intolerant of prolongedantiplatelet therapy and therefore, if the antiplatelet therapy isstopped, they are at risk for thrombosis. Patients treated withdrug-eluting stents are also at greater risk for late stent thrombosisthan patients treated with bare metal stents. These factors areimportant limitations to the uniform use of drug-eluting stents. Thus,the invention provides a method to risk stratify patients undergoingstenting in order to appropriately choose whether a drug-eluting stentis necessary. Based on the risk assessment, those patients above aspecific threshold would receive a DES and may also receive a higherdose of the antiproliferative drug or an alternative antiproliferativedrug(s) as compared to patients at a lower risk threshold.

The present invention is based on the discovery that the maximum tensilestrength of a clot, and more specifically, the maximum clot strength asmeasured by thrombelastography is a powerful marker of the risk ofrestenosis in patients treated with stents for obstructive coronaryartery disease. Importantly, maximum clot strength has been found to bean effective marker of the risk of restenosis irrespective of clinicaland angiographic variables.

Prior to the invention, there was no readily available method to assessrestenosis risk. More importantly, knowledge of the patient's risk forrestenosis is invaluable in preventing complications as those patientsin the highest risk group would be most carefully followed clinically.

Prior to the invention, the drug dose delivered by the drug-elutingstents was uniform; a choice of dose was unavailable. The inventionprovides a scheme for the implementation of various drug doses based onthe patient's risk profile.

In one embodiment, the patient has his/her blood drawn prior to thestent procedure. The blood is analyzed by thrombelastography and themaximum tensile strength of the clot is recorded. The clot usually isstimulated to form by the addition of kaolin, but other agonists thatactivate the generation of thrombin can also be used. The tensilestrength of the given clot is then assessed for restenosis risk based onthe known distribution of tensile strength measured in patients withcoronary artery disease. The patient can then be placed in a risk groupbased on the quartile of clot strength. For example, the 1^(st) quartileis associated with the lowest risk, the 2^(nd) quartile with a higherrisk and so forth up to the 4^(th) quartile where risk is greatest.Based on the quartile of clot strength, the decision for the particularstent can be made. In those patients with 2^(nd)-4^(th) quartile clotstrength, drug-eluting stents should be considered, whereas in thosepatients with the lowest quartile, a bare metal stent would be chosen.

Accordingly, it is a first object of the present invention to detectrestenosis risk in patients receiving a stent by measuring maximumthrombin-induced clot strength.

It is a further object of the present invention to provide such a methodin which the relative risks of respective patients are quantified basedupon four quartiles of clot strength.

It is a yet further object of the present invention to provide such amethod in which those patients falling within the lowest quartile(s)would be treated with bare metal stents.

It is a still further object of the present invention to provide such amethod in which those patients falling within the highest quartile(s)would be treated with a stent coated with a drug-eluting substance.

It is a still further object of the present invention to provide such amethod in which judgments would be made concerning those patientsfalling within the middle 2 quartiles as to whether they are suitablefor a bare metal stent or a stent coated with a drug-eluting substance.

These and other objects, aspects and features of the present inventionwill be better understood from the following detailed description of thepreferred embodiments when read in conjunction with the appended drawingfigures.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1a shows a schematic representation of the structure of a TEGhemostasis analyzer.

FIG. 1b shows a graph of amplitude of the thrombin-generated clot versustime.

FIG. 2 shows a bar graph comparing patients without restenosis withpatients with restenosis including values for maximum amplitude ofplatelet-fibrin mediated clot strength in millimeters (MA) of less than72 and greater than 72, respectively.

SPECIFIC DESCRIPTION OF THE PREFERRED EMBODIMENTS

Platelets play a fundamental role in the genesis of stent restenosis bymediating inflammation and smooth muscle proliferation. Thrombi withhigh tensile strength may facilitate growth factor release to theadjacent vessel wall resulting in neointimal hyperplasia.

Applicant measured pretreatment ex vivo maximum thrombin-induced clotstrength, time to fibrin formation, and the combined clotting index withthrombelastography (TEG) in 178 consecutive patients undergoing electivestenting of de novo (n=160) and restenotic lesions (n=18) and in 25healthy controls. Patients were followed for 6 months for thedevelopment of symptomatic restenosis. Patients who developed restenosis(n=28) had greater maximum clot strength (71.0±5.0 vs. 66.1±7.0,p=0.0004), higher combined clotting indices (3.4±1.8 vs 2.1±2.3,p=0.005) and more rapid fibrin generation times (4.0±1.0 vs 4.8±1.7,p=0.03) than patients without restenosis and healthy controls (p<0.05for all measurements). The relative risk of developing restenosis was22.5 times greater for patients with clot strength in the highestquartile vs. the lowest quartile. Multivariate analysis demonstrated theindependent predictive value of these laboratory measurements.

These results suggest that the prothrombotic state, measured by TEG andhighlighted by increased clot strength, is independently predictive ofthe development of in-stent restenosis. High clot strength may serve asa new marker leading to patient-specific therapies targetingantiproliferative therapy for those at greatest risk.

Presently, platelet related periprocedural thrombotic and inflammatoryprocesses that influence neointimal hyperplasia and angiogenesis areconsidered as important risk factors for restenosis. Despite theseestablished mechanisms, there are few data in humans that examine therelation of ex vivo measurements of platelet reactivity to restenosis.Moreover, preexisting inflammatory and hypercogulable factors are alsosuggested as important factors influencing restenosis. Applicant hasfound that patients with high clot strength may be at greatest risk forrestenosis since their thrombi may be most resistant to disruption byflowing blood. A robust clot would theoretically facilitate residence ofplatelets at the vessel wall and subsequent delivery of pro-restenoticfactors. Moreover, patients with rapid thrombin generation would beexpected to readily form thrombi.

The Investigational Review Board at Sinai Hospital of Baltimore approveda study resulting in development of the present invention. One hundredand seventy-eight consecutive patients who underwent successful electivecoronary artery or saphenous vein graft stenting gave informed consentprior to the procedure and were prospectively followed post-dischargefor the development of symptomatic restenosis. Inclusion criteriaincluded patients over 18 years old. Exclusion criteria were a historyof bleeding diathesis, acute myocardial infarction within 48 hours,elevated cardiac markers (above upper limits normal for the respectiveassay), cerebrovascular event within 3 months, chronic vessel occlusionor angiographically visible thrombus, illicit drug or alcohol abuse,prothrombin time greater than 1.5 times control, plateletcount<100,000/mm³, hematocrit<30%, creatinine>4.0 mg/dl, andglycoprotein (GP) IIb/IIIa inhibitor use prior to the procedure.

Blood samples were obtained in the catheterization laboratory through anindwelling femoral vessel sheath and transferred to vacutainercollecting tubes (Becton-Dickinson, Rutherford, NJ) containing 40 USPlithium heparin after discarding the first 2-3 ml of free flowing blood.Samples were obtained before GPIIb/IIIa inhibitors and heparinadministration (baseline). Blood samples were also obtained at the timeof the second procedure from patients who developed symptomaticrestenosis.

The Thrombelastograph (TEG®) Hemostasis Analyzer with automatedanalytical software provides quantitative and qualitative measurementsof the physical properties of a clot. Briefly, the TEG is a viscoelasticmonitor that measures the degree of platelet-fibrin mediated clotstrength as shown in FIG. 1a . Fibrin strands in the blood sample link arotating sample cup with a stationary pin suspended by a torsion wire.The torque of the rotating cup is transmitted to the immersed pin. Pinmovement is converted to an electrical signal by a transducer and isinterpreted by the computer to create a tracing. The degree of plateletcontribution to the clot strength through platelet-fibrin bondingdirectly influences the magnitude of pin movement and ultimately theamplitude of the tracing. In the present study, the maximum amplitude ofthe thrombin-generated clot [(MA) (mm)], the time from the start of thesample run to the first significant levels of clot formation [reactiontime (R) (min)] and Coagulation index (CI) were measured. Arepresentative signature waveform is shown in FIG. 1b . The resultanthemostasis profile can be evaluated with individual points in theprofile indicating specific parameters of patient hemostasis.

The R parameter is a measure of initial thrombin-generated fibrinformation and has been correlated with the velocity of thrombingeneration. The coagulation index (CI) is a measure of overallcoagulation that is calculated from the kinetics of clot development andformation in native or kaolin-activated whole blood. CI is derived froma computer calculated linear combination of R, K [the rate of thrombusformation], MA and the α-angle [a measure of the rapidity of fibringeneration and cross linking according to the following formula]:

(CI=−0.6516R−0.3772K+0.1224MA+0.0759α−7.7922).

Normal values for the CI lie between −3.0 and +3.0, which is equivalentto three standard deviations about the mean of zero. Positive valuesoutside this range (CI>+3.0) indicate that the sample ishypercoagulable, whereas negative values outside this range (CI<−3.0)indicate that the sample is hypocoagulable.

Blood was analyzed according to the manufacturer's instructions. One mLof heparinized blood was transferred to a vial containing kaolin andmixed by inversion. Five hundred microliters of the activated blood wasthen transferred to a vial containing heparinase and mixed to neutralizethe heparin. The neutralized blood (360 uL) was immediately added to aheparinase coated cup and assayed in the TEG analyzer according to themanufacturer's instructions to obtain the thrombin-induced clot. Oncethe sample assay is completed, automated analytical software generatesthe MA, R and CI values.

Patients were clinically followed by research staff for the developmentof recurrent angina defined as the occurrence of typical symptoms thatwarranted cardiac catheterization at the discretion of the treatingcardiologist who was blinded to the study results of the given patient.Restenosis was defined as >50% luminal diameter stenosis in the stentedsegment by visual estimate. Stent thrombosis was defined by the suddenonset of coronary artery occlusion in a stented vessel resulting inhospitalization and judged by the treating interventionalist as due tothrombosis.

The linear logistic regression model was employed to fit the binary data(restenosis=1 and no-restenosis=0). This logistic regression model wasfit using SAS procedure PROC LOGISTIC (Cary, N.C., USA). The model isgiven by:

Logit(p)=Log{p/(1−p)}=β₀+β₁ ·X

where p=proportion of incidence of restenosis, and X is a predictorvariable. The three variables (possible X) considered here are MA, LTAand R. β₀ is constant or intercept (value of the dependent variable whenX=0); β₁ is the slope (increase in the value of p per unit increase inX).

The multiple linear logistic regression model was employed to fit binarydata to compare the prevalence of demographic, angiographic and clottingvariables in patients with restenosis and without restenosis. Thelogistic regression model was fit using SAS procedure PROC LOGISTIC.Odds ratios were calculated using SAS software, and ROC curves weregenerated using MedCalc Software (MedCalc Software, Broekstraat,Belgium). Based on the normal distribution of data the mean±SD isreported except as otherwise noted and p<0.05 was consideredsignificant. Comparisons were made between the restenotic andnon-restenotic groups by t-tests (Statistica software) for continuousvariables and by Fischer's exact test for categorical data.

The results were as follows:

One hundred thirty-three patients received a loading dose of clopidogrel[300 mg (n=76), 600 mg (n=57)] in the catheterization laboratoryimmediately after successful stenting. Patients on a maintenance dose ofclopidogrel at the time of admission (n=45) did not receive a loadingdose. The GP IIb/IIIa inhibitor, eptifibatide was administered to 71patients at the discretion of the treating physician. All patientstreated with eptifibatide received unfractionated heparin according tothe Enhanced Suppression of the Platelet IIb/IIIa Receptor withIntegrilin Therapy (ESPRIT) dosing regimen (60 U/kg) as a bolus. In allother patients, heparin was administered to achieve an activatedclotting time of 300 seconds. Aspirin (325 mg) was administered on theday of the procedure and daily thereafter. The maintenance dose ofclopidogrel was 75 mg daily.

One hundred and sixty patients underwent stenting of de novo lesions and18 patients underwent stenting of at least one restenotic lesion.Symptomatic restenosis developed in 12 patients over the 6 months offollow-up (10/160 initially treated with de novo lesions land 2/18initially treated with at least one restenotic lesion). 4 patientsdeveloped a myocardial infarction in the distribution of the targetvessel. Among the latter patients, 2 events were due to obstruction ofthe stented segment, and 2 patients had the frank occurrence of asubacute stent thrombosis (Tables 5, 6).

Healthy controls comprised 50% men at an age of 39±8 years and were freeof cardiovascular risk factors and concomitant pharmacologic therapy.The clinical and angiographic demographics of patients with and withoutsymptom driven restenosis are shown in Tables 1 and 2 and demonstrateequivalence between the groups except for a higher incidence of priormyocardial infarction in patients who developed restenosis.

Maximum Clot Strength, Rapidity of Fibrin Formation, and Clot IndexBaseline Measurements

Patients undergoing stenting had markedly higher MA (67.0±6.0) and CI(2.3±2.0); and more rapid fibrin generation (4.7±1.5) than healthycontrols (p<0.001 for all comparisons, Table 3). Patients undergoingtreatment of restenotic lesions had higher baseline MA (69.9±3.0 vs.67.0±6.0, p=0.04) and CI (3.6±1.5 vs. 2.2±2.2, p=0.005), and shorter R(3.7±1.2 vs. 4.8±1.6, p=0.009) than patients undergoing treatment of denovo lesions.

The mean normal value+2SD (72 mm) for MA (MA72) was evaluated todetermine whether patients might be classified by restenosis risk basedon having pre-treatment values above or below this high level ofplatelet-fibrin mediated clot strength (Table 4). Of the 12 patients whosuffered restenosis, 8 had MA values above 72 mm (67%), compared with 30of 150 patients (20%) who did not develop restenosis (p=0.0005). Theodds ratio (OR) of symptomatic restenosis at 6 months post-PCI forpatients with MA values greater than 72 mm was 10.9. Use of MA72 todifferentiate risk of restenosis among post-PCI patients had asensitivity of 67% and a specificity of 80% (FIG. 2). Using the ROCcurves, the sensitivity and specificity are similar to the Table 4results, with a sensitivity of 67% and a specificity of 83%. Eleven outof 12 (92%) patients with restenosis had MA values in the 3^(rd) and4^(th) quartiles (MA>68); eight out of 12 (67%) were in the 4^(th)quartile (MA>72).

Overall, patients who developed restenosis had higher baseline maximumclot strength (75.5.±4.5 vs. 66.5±6.5, p=0.0001), more rapid fibringeneration (4.2±1.0 vs. 4.6±1.5, p=0.36), and a greater clot index(3.6±1.3 vs. 2.3±2.31 p=0.03) than patients who did not developrestenosis. Comparison of measurements with respect to the initialtreatment of de novo vs. restenotic lesions demonstrates that high clotstrength, rapid fibrin generation and a great clot index were associatedwith restenotic lesions. Similar findings occurred in the baselineanalyses from the immediate study that demonstrated higher clotstrength, more rapid fibrin generation and a greater clot index in thosepatients undergoing revascularization for stent restenosis as comparedto treatment of a de nove lesion (Table 3).

The relative risk ratios for restenosis for each of the quartiles ascompared to the first quartile based on the baseline measurements fromboth groups are shown in Table 4.

Discussion

The current study demonstrates that: 1) The physical properties of exvivo thrombi generated by thrombin in the blood of patients undergoingelective stenting differs dramatically from the thrombi generated inhealthy controls. Patient thrombi have markedly greater maximum tensilestrength and fibrin production occurs more rapidly, 2) The physicalproperties of ex vivo thrombi differs between patients undergoingtreatment of de novo lesions versus patients undergoing treatment ofrestenotic lesions. Patients with restenotic lesions have greaterthrombin-induced clot strength and produce fibrin more rapidly thanpatients undergoing treatment of de novo lesions, and 3) Baselinemeasurements of maximum thrombin-induced clot strength are highlypredictive of restenosis; patients in the highest quartile have anapproximate 23 fold risk of developing restenosis as compared topatients in the lowest quartile. Applicant's data demonstrates thatthese measurements are stable over the period of restenosis development.

The current study is the first to examine the predictive value of thephysical properties (i.e. maximum tensile strength) of a patient's clotin determining restenosis. Moreover, it is the first study to assess theutility of thrombelastography measurements as markers for restenosisdevelopment in patients undergoing percutaneous coronaryrevascularization. Moreover, Applicant's data demonstrates theindependent predictive value of the measurements irrespective of theclinical demographics that have been associated with restenosis.

The current study strengthens the proposed central role of reactiveplatelets in the pathophysiology of restenosis. The maximumthrombin-induced clot strength measured by TEG is indicative of thetensile properties of the platelet-fibrin interaction. Applicant hadhypothesized that robust clot formation may facilitate plateletresidence at the vessel wall interface and thus enhance the transfer ofgrowth factors, thus promoting intimal proliferation and restenosis.Platelet and fibrin deposition has been well described at the site ofstent implantation. Platelet surface receptors facilitate attachment ofleukocytes to the area of injury that further release growth factors.Moreover, in the current study, patients with the most rapid thrombingeneration may be expected in vivo to generate activated platelets.Applicant's data supports the importance of the rapidity of thrombingeneration and the responsiveness of platelets to thrombin as majorfactors in the development of restenosis.

It is well known that many patients experience excellent long-termclinical outcomes following treatment with non-drug eluting stents. Inaddition, there are pitfalls to the implantation of drug-eluting stentsthat include a requirement of prolonged dual antiplatelet therapy andconcerns for late stent thrombosis. Prolonged therapy with clopidogreladds significantly to cost and many patients are intolerant totreatment. Up to this date, there have been no validated markers thatpredict restenosis. The validation of a restenosis marker may assist inthe decision making for a bare metal stent as compared to a drug-elutingstent. Applicant's data strongly suggests that patients in the lowestquartile for clot strength are at very low risk of developing restenosiswhereas those in the highest quartile have extremely high risk. Thesequartiles may serve as cutpoints to be examined in prospectiveinvestigations of bare metal versus drug-eluting stents.

Applicant's results suggest that the prothrombotic state measured by TEGand highlighted by increased clot strength is independently predictiveof the development of in-stent restenosis. High clot strength may serveas a new marker leading to patient-specific therapies targetingantiproliferative therapy for those at greatest risk.

As such, an invention has been disclosed in terms of preferredembodiments thereof which fulfill each and every one of the objects ofthe invention as set forth hereinabove, and provide a new and usefulmethod for the detection of restenosis risk in patients receiving astent by measuring the characteristics of blood clotting including themeasurement of maximum thrombin-induced clot strength of great noveltyand utility.

Of course, various changes, alterations and modifications in theteachings of the present invention may be contemplated by those skilledin the art without departing from the intended spirit and scope thereof.As such, it is intended that the present invention only be limited bythe terms of the appended claims.

TABLE 1 Patient Demographics No Restenosis Restenosis (n = 12) (n = 166)p-value Age (years) 66 ± 9  64 ± 12 NS Race (Caucasian) n, (%) 6 (50)121 (73)  .03 Gender (Male) n, (%) 4 (33) 109 (66)   .007 BMI 30 ± 6 30± 7 NS Risk Factors/Past medical Hx n, (%) Smoking 9 (75) 86 (52) .05Family history of CAD 6 (50) 91 (55) NS Hypertension 6 (50) 105 (63)  NSHyperlipidemia 10 (83)  126 (76)  NS Diabetes 5 (42) 73 (44) NS PriorMyocardial Infarction 7 (58) 48 (29)  .009 CHF 3 (25) 24 (14) NS PriorCABG 4 (33) 42 (25) NS Prior PTCA 5 (42) 44 (27) NS Baseline Medicationsn, (%) Aspirin 12 (100) 166 (100) NS Clopidogrel 2 (17) 43 (26) NS Betablockers 8 (67) 147 (89)  .01 ACE Inhibitors 9 (75) 117 (70)  NS Calciumblockers 2 (17) 39 (23) NS Lipid lowering agents 11 (92)  142 (86)  NS3A4 10 (82)  91 (55) NS Non 3A4 1 (8)  51 (31) NS Laboratory Data WBC(×1000/mm³)  7.8 ± 2.6  7.3 ± 2.2 NS Platelets (×1000/mm³) 263 ± 82 236± 69 NS Hematocrit (g/dl) 37.7 ± 5.5 40.0 ± 5.0 NS Hemoglobin (g/dl)12.2 ± 2.3 13.4 ± 1.9 NS Creatinine (g/dl)  1.0 ± 0.3  1.1 ± 0.5 NS BMI= body mass index CABG = coronary artery bypass graft surgery CAD =coronary artery disease PTCA = percutaneous coronary angioplasty ACE =angiotensin converting enzyme WBC = white blood cells 3A4 = hepaticcytochrome 3A4

TABLE 2 Procedural Characteristics No Restenosis Restenosis (n = 12) (n= 166) p Value Ejection Fraction (%) 46 ± 12 50 ± 8  NS Number ofvessels treated 1.3 ± .5 1.3 ± .6  NS Lesion Morphology Denovo n, (%) 10(83)  150 (90)  NS Lesion Location n, (%) LAD 4 (34) 54 (33) NS CX 1 (8)44 (27) .07 RCA 7 (58) 61 (36) .06 SVG 0 7 (4) NS Stent Types Drugeluting n, (%) 8 (66) 116 (70)  NS Reference vessel diameter (mm) 3.1 ±0.4 3.0 ± 0.4 NS Total lesion length (mm) 24 ± 13 19 ± 12 NSPre-stenosis (%) 88  85  NS Post-stenosis (%) 5 4 NS CX = circumflexartery LAD = left anterior descending artery RCA = right coronary arterySVG = saphenous vein graft

TABLE 3 Clot Strength, Reaction Time, Coagulation Index in HealthyControls and Patients with Baseline Denovo and Restenotic LesionsPatients with Baseline De novo Lesions Patients with Baseline RestenoticLesions No Restenosis No Restenosis Restenosis (n = 10) Restenosis (n =2) Healthy (n = 150) At time of (n = 16) At time of Controls BaselineBaseline PCI for Baseline Baseline PCI for (n = 25) MeasurementMeasurement restenosis Measurement Measurement recurrent restenosis ClotStrength (MA) (mm) 60.7 ± 5.1 66.2 ± 7.0*   75.0 ± 4.7** 77.4 ± 4.4 68.5 ± 3.1  76.5 ± 1.0^(Ψ ) 76.6 ± 1.1  Reaction Time (R) (min)  6.9 ±1.2 4.8 ± 1.7* 4.8 ± 1.2 4.6 ± 1.5 4.0 ± 1.1 2.0 ± 0.5^(#) 2.0 ± 0.5Coagulation Index (CI) −0.8 ± 1.8 2.1 ± 2.3* 3.1 ± 1.6 3.8 ± 1.4 3.2 ±1.3 5.9 ± 0.4^(δ) 6.0 ± 0.5 *p < 0.001 Healthy controls vs. patientswith baseline de novo lesions who did not develop restenosis **p < 0.001Patients with baseline de novo lesions who did not develop restenosisvs. who developed restenosis ^(Ψ)p = 0.02 Patients with BaselineRestenotic Lesions who did not develop restenosis vs. who developedrestenosis ^(#)p = 0.002 Patients with Baseline Restenotic Lesions whodid not develop restenosis vs. who developed restenosis ^(δ)p = 0.01Patients with Baseline Restenotic Lesions who did not develop restenosisvs. who developed restenosis

TABLE 4 Relative Risk of Restenosis by Quartiles Relative Risk ofQuartiles Restenosis Clot Strength (MA) 1^(st) 1 2^(nd) 2.5 3^(rd) 54^(th) 22.5 Reaction Time (R) 1^(st) 1 2^(nd) 3 3^(rd) 4 4^(th) 2Coagulation Index (CI) 1^(st) 1 2^(nd) 3 3^(rd) 3 4^(th) 7

TABLE 5 Comparison of sensitivity and specificity of MA72 in predictingrestenosis at 6 months post-PCI Restenosis No Restenosis (n) (n) Total(n) Odds Ratio MA > 72 mm 8  30  38 10.9 MA ≤ 72 mm 4 120 124 — Total 12150 162 — Sensitivity (%) 67 — — — Specificity (%) 80 — — —

TABLE 6 Stepwise multiple logistic regression analysis Estimate SE pvalue OR 95% CI Intercept −2.25 1.87 .2305 — — Gender 2.11 0.85 .01298.26  1.57, 43.64 Prior PTCA −1.43 0.72 .0478 0.24 .06, .99 Prior MI−1.54 0.80 .0542 0.21 0.05, 1.03 MA72 2.39 0.79 .0025 10.88  2.31, 51.19No additional effects met the .05 significance level for inclusion inthe model

1. A method of selecting a stent for implantation in the circulatorysystem of a human being including the steps of: a) determining athreshold level of platelet hyper-coaguability (PHC) of blood, abovewhich a risk of restenosis is relatively high; b) obtaining a bloodsample from a patient who requires implantation of a stent; c) testingsaid blood sample for its PHC; d) comparing PHC of said blood samplewith said threshold level; e) if said blood sample has a PHC below saidthreshold level, selecting a bare metal stent; and f) if said bloodsample has a PHC at or above said threshold level, selecting adrug-eluting stent.
 2. The method of claim 1, wherein said determiningstep includes the step of obtaining blood samples from a multiplicity ofpeople, testing each sample for PHC and correlating said samples withdata regarding onset of restenosis among said people to determine saidthreshold level.
 3. The method of claim 1, wherein a marker of PHCmeasured comprises platelet-fibrin mediated clot strength (MA).
 4. Themethod of claim 3, wherein said threshold level is MA=72.
 5. The methodof claim 3, wherein said testing step comprises: a) obtaining aviscoelastic monitor including a blood receiving cup; b) pouring bloodinto said cup; c) oscillating said cup with a pin suspended in saidblood by a torsion wire; d) when clotting commences, said cup and pinbecome interconnected; e) measuring clot strength.
 6. The method ofclaim 5, wherein said viscoelastic monitor includes automated analyticalsoftware.
 7. The method of claim 6, wherein said software providesquantitative and qualitative measurements of initial fibrin formation,kinetics of clotting process, degree of platelet-fibrin mediated clotstrength (MA), and dissolution.
 8. The method of claim 5, wherein saidoscillating step occurs over a distance of 4° 45′.
 9. The method ofclaim 8, wherein said oscillating step occurs in 10 second cycles. 10.The method of claim 5, wherein said blood is mixed with a clot-promotingsub stance.
 11. The method of claim 10, wherein said substance compriseskaolin.
 12. The method of claim 3, wherein results from testing saidsamples are divided into quartiles sequentially numbered 1-4 from lowestto highest MA.
 13. The method of claim 12, wherein patients having an MAin a 4^(th) said quartile are implanted with a drug-eluting stent. 14.The method of claim 12, wherein patients having an MA in a 1^(st) saidquartile are implanted with a bare metal stent.
 15. The method of claim1, wherein a marker of PHC measured comprises speed of thrombingeneration.
 16. The method of claim 1, wherein a marker of PHC measuredcomprises platelet reactivity.
 17. The method of claim 16, wherein saidplatelet reactivity is measured by an aggregometer.
 18. The method ofclaim 16, wherein said platelet reactivity is measured by flowcytometry.
 19. A method of selecting a stent for implantation in thecirculatory system of a human being including the steps of: a)determining a threshold level of platelet-fibrin mediated clot strength(MA) of blood, above which a risk of restenosis is relatively high; b)obtaining a blood sample from a patient who requires implantation of astent; c) testing said blood sample for its platelet-fibrin mediatedclot strength, wherein said testing step comprises: i) obtaining aviscoelastic monitor including a blood receiving cup; ii) pouring bloodinto said cup; iii) oscillating said cup with a pin suspended in saidblood by a torsion wire; iv) when clotting commences, said cup and pinbecome interconnected; v) measuring clot strength; vi) said viscoelasticmonitor including automated analytical software; d) comparing clotstrength of said blood sample with said threshold level; e) if saidblood sample has a clot strength below said threshold level, selecting abare metal stent; and f) if said blood sample has a clot strength at orabove said threshold level, selecting a drug-eluting stent.
 20. Themethod of claim 19, wherein said step includes the step of obtainingblood samples from a multiplicity of people, testing each sample forplatelet-fibrin mediated clot strength and correlating said samples withdata regarding onset of restenosis among said people to determine saidthreshold level.
 21. The method of claim 20, wherein said thresholdlevel is MA=72.
 22. The method of claim 19, wherein said softwareprovides quantitative and qualitative measurements of initial fibrinformation, kinetics of clotting process, degree of platelet-fibrinmediated clot strength (MA), and dissolution.
 23. The method of claim19, wherein said oscillating step occurs over a distance of 4° 15′ andin 10 second cycles.
 24. The method of claim 19, wherein said blood ismixed with a clot-promoting substance comprising kaolin.